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1、應(yīng)用與環(huán)境生物學(xué)報2010，16(2):274~2782010-04-25ChinJApplEnvironBiol=ISSN1006-687XDOI:10.3724/SP.J.1145.2010.00274纖維素酶產(chǎn)生菌HS-F9的篩選鑒定和產(chǎn)酶條件優(yōu)化*魏艷紅1熊鷹1袁永澤1鄭揚忠1胡柳如1張華1李風(fēng)1楊江科2閆云君2劉德立1**1（華中師范大學(xué)生命科學(xué)學(xué)院遺傳調(diào)控與整合生物學(xué)重點實驗室武漢430079）2（華中科技大學(xué)生命科學(xué)與技術(shù)學(xué)院武漢430074）摘要為了選育高產(chǎn)纖維素酶菌株，通過剛果紅鑒別培養(yǎng)基以及
2、濾紙條崩解實驗測定，從牛糞堆肥中篩選到一株產(chǎn)纖維素酶的真菌HS-F9，根據(jù)菌株形態(tài)特性和18SrRNA基因序列分析，初步鑒定該菌為綠色木霉（Trichodermaviride）.利用液體發(fā)酵培養(yǎng)產(chǎn)生纖維素酶，研究了碳源、氮源、培養(yǎng)時間、培養(yǎng)溫度、培養(yǎng)基起始pH、接種量對菌株HS-F9產(chǎn)酶的影響.結(jié)果表明：產(chǎn)EG、CBH和FPA的最適碳源均為CMC-Na；EG和CBH在以蛋白胨為唯一氮源時酶活最高，F(xiàn)Pa?e則在以黃豆粉為唯一氮源時酶活最高；產(chǎn)生EG、CBH和FPA的最適溫度分別為30℃、30℃、33℃；最適起始
3、pH為3.0、3.0和4.0；EG和FPa?e的最適接種量為2%，CBH最適接種量達到了8%；培養(yǎng)時間均以5~6?為宜.在最適條件下培養(yǎng)，該菌株EG、CBH和FPa?e的酶活分別達到了5275.3U/??L、8502.1U/??L和3619.1U/??L，是未優(yōu)化前的1.42、1.35和1.32倍.圖6表2參23關(guān)鍵詞纖維素酶；菌株篩選；產(chǎn)酶條件優(yōu)化；酶活性CLCTQ925:Q936ScreeningandIdentificationofaCellulase-producingFungusandOptimiza
4、tionofItsFermentationConditions*WEIYanhong1,XIONGYing1,YUANYongze1,ZHENGYangzhong1,HULiuru1,ZHANGHua1,LIFeng1,YANGJangke2,YANYunjun2&LIUDeli1**(1LaboratoryofGeneticRegulationandIntegrativeBiology,CollegeofLifeSciences,HuazhongNormalUniversity,Wuhan430079,Chi
5、na)(2CollegeofLifeSciencesandTechnology,HuazhongUniversityofScienceandTechnology,Wuhan430049,China)AbstractInor?ertoerto?reeree?high-?high-?ieliel???traintrain???orcellula?e,ae,a?unguungu?(HS-F9)?(HS-F9)?ithhighcellula?eacti?itit???a?ia?i?olate?olate??ro??co
6、co??dungthroughcongoredidentificationmediumandthecollapseoffilter,andwasidentifiedasTrichodermaviride??anal?zingitsmorphologyand18SrRNAgenesequences.Differentconditionsincludingcarbonsource,nitrogensource,incubatingtime,growthtemperature,initialpHandinoculat
7、ionquantityfortheproductionofcellulasewerestudiedwithliquid-statefermentation.TheoptimumculturalconditionsofstrainHS-F9forthecellulaseproductionwereasfollows:TheoptimumcarbonsourceforproducingEG(endo-β-1,4-glucana?e?),CBH(exo-β-1,4-glucanasesorcellobiohy-dro
8、lase)andFPase(filterpaperenzyme)wasCMC-Na;theoptimumnitrogensourcewaspeptoneforEGandCBH,butsoyflourforFPase;theoptimumtemperaturesforEG,CBHan?FPa?e?ere30℃,30℃an?33℃,andtheoptimuminitialpHvaluesw
